Particulate Science and Technology

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A novel green synthesis of gold nanoparticles

using seaweed Lobophora variegata and its
potential application in the reduction of
nitrophenols

Princy Kaithavelikkakath Francis, Sreeja Sivadasan, Anna Avarachan & Anu

Gopinath

To cite this article: Princy Kaithavelikkakath Francis, Sreeja Sivadasan, Anna Avarachan &
Anu Gopinath (2019): A novel green synthesis of gold nanoparticles using seaweed Lobophora
variegata and its potential application in the reduction of nitrophenols, Particulate Science and
Technology, DOI: 10.1080/02726351.2018.1547340

To link to this article: https://doi.org/10.1080/02726351.2018.1547340

Published online: 03 Jan 2019.

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PARTICULATE SCIENCE AND TECHNOLOGY
https://doi.org/10.1080/02726351.2018.1547340

A novel green synthesis of gold nanoparticles using seaweed Lobophora

variegata and its potential application in the reduction of nitrophenols
Princy Kaithavelikkakath Francisa, Sreeja Sivadasanb, Anna Avarachana, and Anu Gopinathc
a
Department of Chemistry, St. Albert’s College, Ernakulam, India; bDepartment of Marine Chemistry, Kerala University of Fisheries and Ocean
Studies, Cochin, India; cDepartment of Fishery Hydrography, Kerala University of Fisheries and Ocean Studies, Cochin, India

ABSTRACT KEYWORDS
Developing a facile and green approach to synthesize gold nanoparticles (AuNPs) with high catalytic Green synthesis; gold
activities for the reduction of toxic nitrophenols to beneficial aminophenols is of tremendous envir- nanoparticles; Lobophora
onmental and industrial relevance. The present study reports an environmentally benign synthesis variegata; nanocatalyst;
Nitrophenols
of AuNPs using aqueous extract of seaweed Lobophora variegata as a potential bioreductant. The
signatory pinkish red color and a surface Plasmon resonance peak at 530 nm in the UV-Visible spec-
trum confirmed the formation of AuNPs. TEM images revealed that the biosynthesised AuNPs were
spherical in shape with an average size of 11.69 þ 2.38 nm. The high crystallinity of the biogenic
AuNPs is confirmed from the clear lattice fringes in the High resolution TEM images, bright circular
rings in SAED pattern and Bragg’s reflections of (111), (200), and (220) in XRD pattern. FTIR spectra
revealed the role of bioactive metabolites in stabilizing the nanoparticles. The report emphasizes
the spectroscopic investigation on the efficacy of the biosynthesized AuNPs as excellent catalyst in
the reduction of anthropogenic organic pollutants, 4-nitrophenol (4NP), 3-nitrophenol (3NP), and
2-nitrophenol (2NP) to the corresponding aminophenols using NaBH4, which followed a pseudo-
first-order kinetics with rate constants in the order k2NP > k4NP > k3NP.

1. Introduction
Nitrophenols (NP) are anthropogenic organic pollutants in
wastewater produced from industrial plants for the manufac-
ture of pesticides, insecticides, herbicides, explosives, and
synthetic dyes (Higson 1992). Because of the carcinogenic
and mutagenic properties, these are very harmful to human
beings and aquatic organisms. They are enrolled as a toxin
by the United States Environment Protection Agency
(Arora, Srivastava, and Singh 2014; Pandey and Mishra
2014). Conversely, its reduction product aminophenols (AP)
have a wide range of applications such as precursors for the
manufacture of several analgesic and antipyretic drugs, as
photographic developer, as anticorrosion agent in paints,
and in dye industries (Vaidya, Kulkarni, and Chaudhari
2003; Nemanashi and Meijboom 2013). One of the promin-
ent methods to convert toxic nitrophenols to beneficial ami-
nophenols is to carry out their reduction using NaBH4.
However, this reaction is insignificantly slow in the absence
of a catalyst. In recent times, nanocatalysis has emerged as a
rapidly developing area of research in which metal nanopar-
ticles are used as catalysts for wide range of chemical reac-
tions. Small size, high surface area to volume fraction, and
size relative activity have made metal nanoparticle an effi-
cient catalyst. The potential catalytic ability of noble metal
nanoparticles in the reduction of NP have been reported
recently (Sen, Maity, and Islam 2013; Zhao et al. 2015).
Among noble metal nanoparticles, gold nanoparticles
(AuNPs) are of specific interest owing to their remarkable
surface plasmon resonance (SPR) in the visible region with
extensive applications in various fields such as drug delivery,
catalysis, optoelectronics, surface-enhanced Raman scatter-
ing, biological labeling, bio-imaging, and antimicrobial (Jain
et al. 2008; Kumar et al. 2011). Various physical and chem-
ical methods are available for the synthesis of AuNPs.
However, they are often costly and involve the utilization of
hazardous chemicals that are potentially harmful to the
environment (Menon, Rajeshkumar, and Venkat Kumar
2017; Khandanlou et al. 2018). So, there is a growing inter-
est associated with the development of cost-effective, non-
toxic, and environment friendly methods for the synthesis
of AuNPs.
Nowadays, bio-inspired methods have aroused much
interest due to its simplicity, environment benign nature,
and cost-effectiveness. Biological resources like microbes and
plant extracts were used for this purpose. Among these
methods, the plant-mediated synthesis achieved fabulous
attractiveness due to its eco-friendliness and simplicity.
Moreover, it does not require the complicated process of
maintaining cell culture as in microbe assisted synthesis.
There are numerous reports available for the biosynthesis of
AuNPs using several terrestrial plants (Narayanan and
Sakthivel 2008; Iravani 2011; Philip et al. 2011). However,

CONTACT Anu Gopinath anugowri@hotmail.com Department of Fishery Hydrography, Kerala University of Fisheries and Ocean Studies, Cochin, India.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/upst.
ß 2018 Taylor & Francis Group, LLC
2 P. KAITHAVELIKKAKATH FRANCIS ET AL.

renewable marine resources like marine seaweeds are not

much explored for gold nanosynthesis.
In this work, we report for the first time a novel environ-
mentally benign method for the synthesis of AuNPs using
aqueous extract of brown seaweed Lobophora variegata. The
seaweed extract act as both reducing and capping agent for
the synthesis of AuNPs. Since no toxic reagents used in the
synthetic route, it is completely a green chemistry approach.
Furthermore, this method is facile, fast, and cost-effective.
The catalytic efficacy of the biosynthesized AuNPs was also
studied spectroscopically in the reduction of anthropogenic
organic pollutants, 4-nitrophenol (4NP), 3-nitrophenol
(3NP), and 2-nitrophenol (2NP) to the corresponding AP by
NaBH4 at room temperature in aqueous medium.
2. Materials and methods
2.1. Materials
Chloroauric acid (99.9%) was purchased from Sigma-Aldrich.
4-Nitrophenol, 3- Nitrophenol, 2-nitrophenol, and NaBH4
were procured from Merck India Ltd. All these chemicals were
used as such without further refinements and its aqueous solu-
tions were prepared using sterile Milli Q water.
2.2. Preparation of seaweeds extract
The seaweed L. variegata (Figure 1) was harvested manually
from the coast of Thirumullavaram (8 530 3800 N; 76 330 1400 E),
Kollam district of Kerala and washed it many times in seawater
to remove extraneous materials and then brought to the
laboratory in an ice box. It is then washed with Milli Q water
and air dried. Ten grams of the seaweed was finely chopped
into small pieces and stirred with 100 mL sterile Milli Q water
for 1 min and kept in a water bath at 60  C for 20 min. It was
then cooled and filtered using Whatman No. 1 filter paper,
and the filtrate was used for nanosynthesis.
2.3. Biosynthesis of AuNPs
AuNPs were prepared by adding 0.5 mL of the seaweed extract
to 10 mL of 1 mM Chloroauric acid solution at room tempera-
ture. A control setup was also retained without seaweed
extract. In order to evaluate the effect of quantity of seaweed
extract on gold nanosynthesis, the biosynthetic procedure was
repeated by varying the volume of the seaweed extract
(0.25 mL and 1 mL). The biotransformation of chloroaurate
ions to AuNPs was identified visually by the instant color
change of the reaction medium and was confirmed by UV-Vis.
spectral analysis. The biosynthesized nanoparticles were sepa-
rated by centrifugation at 15,000 rpm for 20 min at 4  C and
purified by redispersing them in Milli Q water. The pure sam-
ples obtained were freeze-dried and kept for analysis.
2.4. Characterization techniques
Thermo scientific Evolution 201 UV–Visible spectrophotom-
eter operating at a resolution of 1 nm in the wavelength
Figure 1. Photograph of seaweed Lobophora variegata.
range of 300–700 nm was used for the UV-vis. spectral stud-
ies. FTIR spectrum of freeze-dried seaweed extract and
AuNPs were recorded using a Perkin Elmer Spectrum 100
spectrometer at the wavelength range of 4000–550 cm1.
The X-ray diffraction (XRD) measurements of drop-coated
films of AuNPs on glass slides were made on a PANalytic
X’PERT-PRO X-ray spectrometer operated at a voltage of
40 kV and a current of 30 mA with Cu Ka radiation
(k ¼ 0.1542nm). High resolution Transmission electron
microscopy (HRTEM) images, Energy dispersive X-ray ana-
lysis (EDAX), and Selected area electron diffraction (SAED)
patterns were taken in Tecnai G2 30 transmission elec-
tron microscope.
2.5. Catalytic activity
The catalytic efficacy of the biosynthesized AuNPs to reduce
the isomers of NP to the corresponding AP by NaBH4 was
evaluated using the method of Gangula et al. (2011), with
slight modification. For this, in a quartz cuvette of 1 cm
path length, 1.4 mL of water, 0.3 mL of 2 mM solution of
NP, and 1 mL of 0.03 M of freshly prepared NaBH4 solutions
were taken. To this reaction mixture, 100 mL of biogenic
AuNPs (1 mg/mL) was added and reduction of the NP was
monitored spectroscopically at regular intervals of time.
3. Results and discussion
3.1. Visual observation and UV-Visible analysis
When L. variegata extract is added to HAuCl4 solution, we
can have a visual perception of change in color of reaction
medium from light yellow to dark pink indicating the for-
mation of AuNPs (Figure 2). The appearance of characteris-
tic pink color was due to the excitation of surface plasmon
resonance (SPR) in AuNPs (Mulvaney, 1996). Moreover,
UV-Visible spectroscopy gave a clear indication of the for-
mation of AuNPs. The UV-Visible spectra depicted in
Figure 3 elucidate the impact of different volumes of sea-
weed extract on the biosynthesis of AuNPs. From the spec-
tra, it is apparent that when the quantity of seaweed extract
is augmented from 0.25 mL to 0.5 mL, the SPR band shifted

Figure 2. Digital images of (a) Lobophora variegata Extract (b) HAuCl4, and
(c) AuNPs.
Figure 3. The UV–Visible absorption spectrum of AuNPs biosynthesized using
varying quantities of Lobophora variegata extract.
towards the shorter wavelength region from 538 nm to
530 nm which shows a decline in particle dimension (Barai
et al. 2018; Princy and Gopinath 2018). But, when the quan-
tity of extract is increased to 1 mL, the SPR band slightly
broadened and showed a red shift to 536 nm indicating an
increase in particle size. Thus, the quantity of extract is a
vital factor in determining the size distribution of AuNPs.
The sharp and symmetric SPR band at 530 nm reflected the
spherical shape of the biosynthesized AuNPs which was fur-
ther set out in TEM analyses. The result from UV-Visible
absorption is in accordance with the previous report (Rajan,
MeenaKumari, and Philip 2014).
Morphological and compositional studies
Transmission electron microscopy is a powerful tool to elu-
cidate the size, shape, and morphology of the nanoparticles.
The TEM images at different magnifications (Figure 4a–d)
proved that the biosynthesized AuNPs possess more or less
spherical geometry. The well-separated particles observed in
TEM images proved the vital role of phytochemicals in
L. variegata extract in capping and stabilizing the nanopar-
ticles. From the histogram representing the size distribution
PARTICULATE SCIENCE AND TECHNOLOGY 3
of the particles (Figure 4f), the average size of the AuNPs
was found to be 11.69 þ 2.38 nm. The clear lattice fringes in
the HR-TEM image of a single nanoparticle (Figure 4d) and
the bright circular rings in the SAED pattern offered clear-
cut evidence to the crystalline nature biosynthesized AuNPs.
The EDAX spectrum of biologically synthesized AuNPs is
depicted in Figure 4g. This spectrum gave a strong peak at
around 2 keV which is due to the presence of metallic gold.
The extra peaks of C and Cu in the spectrum are due to
carbon-coated copper grid used in the TEM analysis.
3.2. Structural analysis
The XRD profile of biosynthesized AuNPs is shown in
Figure 5. The x-ray diffraction peaks appeared at 2h values
of 38.40 , 44.36 , and 64.87 , which can be indexed as
(1 1 1), (2 0 0), and (2 2 0) crystallographic planes respect-
ively, indicating that the green synthesized AuNPs are crys-
talline in nature with face-centered cubic structure (JCPDS
file No. 04-0784). Broad peaks confirmed the formation of
nanosized particles. No other impurity peaks were observed,
which revealed the purity of the AuNPs synthesized.
Further, the peak corresponding to the (1 1 1) plane is more
intense than the other peaks suggesting that predominant
growth of AuNPs oriented along the (1 1 1) plane. The aver-
age crystalline size of the biosynthesized AuNPs was also
determined using Debye-Scherrer equation, by considering
the width of (1 1 1) peak and found to be 11.8 nm which
agrees with the particle size obtained from the TEM image.
3.3. FTIR spectral analysis
FTIR spectroscopy is used to analyze the functional groups
of seaweed extract in capping the nanoparticles. The FTIR
spectrum of the freeze-dried seaweed extract and biosynthe-
sized AuNPs are shown in Figure 6. It is seen that all prom-
inent peaks in the seaweed extract were present in the
AuNPs, which disclose the role of seaweed extract in reduc-
ing and capping the AuNPs. The strong intense broadband
appeared between 3400 and 3450 cm1is due to the stretch-
ing vibrations of O–H groups present in alcohols and phe-
nols (Jilie and Shaoning 2007). A weak peak at 2928 cm1
could be assigned to the C–H stretching vibrations (Suman
et al. 2014; Vijayan, Joseph, and Mathew 2018). Another
intense peak at 1624 cm1 may be due to the –C ¼ O group
coming from the carbonyl group of proteins (MeenaKumari
and Philip 2015). The band located at 1385 cm1 is attrib-
uted to the C–N stretching vibrations (Gavade et al. 2015;
Sadeghi and Gholamhoseinpoor 2015). The absorption peak
at 1028 cm1 corresponds to the carboxylic acid functional
group (Song, Jang, and Kim 2009). These results suggested
that different bioconstituents of seaweed extract have
involved in the reduction as well as stabilization of AuNPs.
But the precise nature and chemical composition of com-
pounds need a detailed investigation.
4 P. KAITHAVELIKKAKATH FRANCIS ET AL.
Figure 4. (a–d) TEM images of AuNPs at different magnifications (e) SAED pattern (f) Particle size histogram, and (g) EDAX spectrum of AuNPs.
Figure 5. XRD pattern of biosynthesized AuNPs.
3.4. Catalytic activity of AuNPs for the reduction of
nitrophenols
Catalytic efficacy of biosynthesized AuNPs was demonstrated
by considering the reduction reactions of three isomers of
nitrophenols, i.e., 4NP, 3NP, and 2NP using NaBH4. For 4NP
(Figure 7a), upon addition of NaBH4, the absorption maxima
underwent a redshift from 316 nm to 400 nm and the yellow
color of the solution become more intense due to the formation
of 4-nitrophenolate ion in the alkaline condition caused by the
NaBH4 and was highly stabilized by resonance (Pradhan, Pal,
and Pal 2001). In the absence of the catalyst, the intensity of the
peak at 400 nm as well as the color of the solution remained
unchanged for several hours, suggesting that the reduction is
insignificantly slow due to the appreciable difference in redox
potential between them (Majumdar, Bag, and Ghosh 2016).
But, in the addition of 100 mL AuNPs (1 mg/mL), reduction
reaction started immediately. This is evidenced from the fading
of yellow color of the reaction medium as well as decrease in
the intensity of the peak at 400 nm. As the intensity of the peak
at 400 nm decreases, a new absorption peak appeared at 300 nm
appeared and its intensity increases with elapse of time. This
new peak is due to the formation of 4-aminophenol, the prod-
uct of the reaction. The reaction was found to be completed in
4 min as was evident from complete bleaching of yellow color
of the reaction mixture as well as by the disappearance of the
absorption peak at 400 nm. Similarly, 3NP and 2NP exhibit
absorption peak at 392 nm and 415 nm, respectively, in the
alkaline medium due to the formation of their corresponding
nitrophenolate ions, both of which decreased readily on the
addition of AuNPs in company with the appearance of new
peaks at 302 nm and 298 nm due to the formation of 3AP and
2AP, respectively (Figure 7b,c). The reduction of 3NP com-
pletes in 8 min while it takes only 3 min for 2NP.
Here, the concentration of NaBH4 (0.03 M) used was very
high compared to NP (2 mM) and remains practically con-
stant throughout the course of the reaction, so the reaction
kinetics is assumed to follow pseudo first order and the rate
equation can be written as
kt ¼ ln ðC=C0 Þ;
where k is the pseudo first-order rate constant, t is reaction
time, and C and C0 are concentration of the NP at time t
and 0, respectively. As the absorbance is proportional to the
concentration of solution, the absorbance at time t (A) and
time 0 (A0) are equivalent to the concentration at time t (C)
and time 0 (C0), respectively (Bindhu and Umadevi 2015).
The linear plots of ln (A/A0) against time in all the three
reactions support the pseudo-first order assumption (inset of
Figure 7a–c). The rate constants estimated from the slopes of
the linear sections of the plots and the corresponding correl-
ation coefficients are listed in Table 1. The observed results are
in consistent with the values reported for Mimusops elengi bark
 PARTICULATE SCIENCE AND TECHNOLOGY 5

Figure 6. FTIR spectrum of (a) Lobophora variegata Extract and (b) biosynthesized AuNPs.

Figure 7. (a) UV-Visible absorption spectra for the NaBH4 reduction of (a) 4 NP, (b) 2NP, and (c) 3NP using 100 mL AuNPs (1 mg/mL). Inset shows the respective
linear plots of ln (A/Ao) versus reaction time.

Table 1. Catalytic activity of 100 mL AuNPs (1 mg/mL) in the reduction of NP.

extract stabilized AuNPs (Majumdar, Bag, and Ghosh 2016). As
NP Reaction time (min) K  103 (sec1) Correlation coefficient (R2)
is evident from the table, all these reactions are very fast and
4NP 4 8.8 0.999
they all strictly hold on to pseudo first-order kinetics as evi- 3NP 8 4.5 0.990
denced from the correlation coefficient values. The rate con- 2NP 3 12.1 0.993
stants (k) of the reactions are estimated to be 8.8  103 sec1,
4.5  103 sec1, and 12.1  103 sec1 for 4NP, 3NP, and 2-
NP respectively. The rates of reduction of three isomers of donor (BH4) and electron acceptor (nitrophenolate ion) get
nitrophenol followed the order 2NP > 4NP > 3NP. adsorbed on the surface of the AuNPs, catalytic reaction
In all these reactions, the enhancement of reaction rate is happens by the transfer of electron from BH4 to nitrophe-
due to the electron relay effect. The AuNPs takes the role of nolate ion. Thus, in all the above catalytic reactions, AuNPs
an electron transfer mediator between the electron donor help in facilitating the reduction by lowering the activation
NaBH4 and the accepter NP. The catalytic reduction takes energy of the reaction and hence play the role of an effective
place on the surface of the AuNPs. As soon as the electron catalyst (Aromal and Philip 2012).
6 P. KAITHAVELIKKAKATH FRANCIS ET AL.
4. Conclusions
In this study, the highly stable and monodispersed AuNPs were
synthesized in a green way using aqueous extract of brown sea-
weed L. variegata as the bioreductant. The seaweed extract also
act as the capping agent for the formed nanoparticles. The vari-
ous characteristics of the biosynthesized AuNPs were analyzed
using UV-Vis, TEM, SAED, EDAX, XRD, and FTIR techni-
ques. The spherical nanoparticles with an average size of
11.69 þ 2.38 nm were formed which exhibited an SPR band at
530 nm in the UV-Vis spectrum. The crystalline nature of the
nanoparticles was evidenced from XRD and SAED patterns.
The excellent catalytic efficacy of the biosynthesized AuNPs
was demonstrated using the NaBH4 reduction of isomers of NP
to the corresponding AP. These reactions were very fast and fol-
low pseudo-first order kinetics. The rate constants of the reac-
tions followed the order 2NP > 4NP > 3NP. Thus, the
biosynthesized AuNPs can act as effective green catalyst for
environmental protection.
Acknowledgments
The authors are delighted to acknowledge STIC, KOCHI, School of Pure
and Applied Physics, M.G. University and NIIST, Thiruvananthapuram
for the technical aid in characterization studies.
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